TruSeq Total RNA Training
15044069
TruSeq Stranded Total RNA provides a clear view of the transcriptome with a streamlined, cost-efficient, and scalable solution for total RNA analysis with sequencing.
Compatible with a wide range of samples, including low-quality DNA/RNA and formalin-fixed, paraffin-embedded (FFPE) samples, TruSeq Stranded Total RNA couples all of the benefits of TruSeq RNA technology with Ribo-Zero ribosomal RNA reduction chemistry. The product enables analysis of coding and multiple forms of noncoding RNA with precise measurement of strand orientation, uniform coverage, and high-confidence discovery of features such as alternative transcripts, gene fusions, and allele-specific expression.
TruSeq Stranded Total RNA is designed for human, mouse and rat sequences, but may be applicable to a variety of eukaryotic species.
TruSeq Stranded Total RNA is available in multiple versions to fit specific study design needs:
| Instrument | Recommended Number of Samples | Read Length |
|---|---|---|
| NextSeq 550 System | RNA Profiling: 6-20 samples per run (based on 20 million reads per
sample) Transcriptome Analysis: 2-8 samples per run (based on 50 million reads per sample) |
2 x 75 bp |
| HiSeq 2500 System | RNA Profiling: 30-200 samples per run (dual flow cell; based on 20
million reads per sample) Transcriptome Analysis: 12-80 samples per run (dual flow cell; based on 50 million reads per sample) |
2 x 75 bp |
| NovaSeq 6000 System | RNA Profiling (samples per run, dual flow cell): S1: 160, S2: 320, S4:
768. Limited by index combinations (dual). Based on 20 million reads. Transcriptome Analysis (samples per run, dual flow cell): S1: 64, S2: 128, S4: 400. Limited by index combinations (dual). Based on 50 million reads. |
≤ 2 × 100 bp |
| TruSeq Stranded Total RNA | Illumina Stranded Total RNA Prep with Ribo-Zero Plus | |
|---|---|---|
| Assay Time | 11.5 hours | ~7 hours |
| Automation Capability | Liquid Handling Robots | Liquid Handling Robots |
| Content Specifications | Captures coding RNA plus multiple forms of non-coding RNA | Captures coding RNA plus multiple forms of non-coding RNA |
| Description | For whole-transcriptome sequencing studies. The Total RNA version depletes cytoplasmic rRNA while the Total RNA Gold version depletes cytoplasmic and mitochondrial rRNA. | For whole-transcriptome sequencing studies of multiple sample types. Includes Ribo-Zero plus module for single-tube depletion of abundant transcripts from multiple species |
| Hands-On Time | 5.5 hours | < 3 hours |
| Input Quantity | 0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization. | 1-1000 ng total RNA for standard-quality RNA samples. 10ng total RNA minimum recommended for optimal performance and FFPE samples |
| Mechanism of Action | Bead-based rRNA depletion, cDNA synthesis, and PCR | Enzymatic rRNA depletion, ligation-based addition of adapters and indexes |
| Method | Whole-Transcriptome Sequencing | Whole-Transcriptome Sequencing |
| Multiplexing | Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together, High-throughput kits: Prepare 96 indexes samples using either combinatorial dual indexes or unique dual indexes (available soon) | Up to 384 Unique Dual Indexes (UDIs) |
| Specialized Sample Types | FFPE Tissue | FFPE Tissue, Low-Input Samples |
| Species Category | Human, Mouse, Rat | Bacteria, Human, Mouse, Rat, Virus |
| Species Details | rRNA Removal: Depletes human, mouse, and rat rRNA | Ribo-Zero Plus included for depletion of abundant transcripts from multiple species including: human cytoplasmic & mitochondria rRNA, mouse rRNA, rat rRNA, bacteria Gram +/- rRNA, human beta globin transcripts |
| Technology | Sequencing | Sequencing |
The products previously known as TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Human/Mouse/Rat (Cat. No. RS-122-2201, RS-122-2202, and RS-122-2203) and TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Cat. No. RS-122-2301, RS-122-2302, and RS-122-2303) are now called TruSeq Stranded Total RNA Human/Mouse/Rat and TruSeq Stranded Total RNA Gold, respectively. The product configuration has changed. In the new configuration, the library preparation reagents and index adapters can be purchased separately. The list of components available for individual purchase can be found in the “Select Product(s)” section.
BaseSpace Sequence Hub helps the Biotechnology Resource Center Sequencing Core (BRC-Seq) at the University of British Columbia deliver high-quality sequencing data and additional value to its customers.
Read More
Consistent, precise measurement of RNA abundance is reflected by high reproducibility between technical replicates. These technical replicates of FFPE tissue show high concordance, indicating robust library prep performance. Axes are log2(FPKM). R2 value is shown.
TruSeq Stranded Total RNA gives excellent coverage across the top 1000 expressed transcripts in both fresh-frozen (FF, top) and FFPE (bottom) tumor and matched normal breast tissue, with > 98% aligned stranded reads. X-axis: position along transcript, Y-axis = percent coverge of combined reads.
Robust low-input RNA sequencing with TruSeq Stranded Total RNA Library Prep Gold
Application Note | HTML
Data Sheet | PDF | 5 versions
TruSeq Stranded Total RNA With Illumina Ribo-Zero Plus rRNA Depletion Reference Guide Documentation
TruSeq Stranded Total RNA Support - Documentation & Literature
TruSeq Stranded Total RNA Consumables & Equipment Documentation
TruSeq Stranded Total RNA Reference Guide Documentation
TruSeq Stranded Total RNA Checklist Documentation
TruSeq Stranded Total RNA Documentation
Custom Protocol Selector
Generates customized, end-to-end instructions
Determine the best library prep kit or array for your needs based on your starting material and method of interest.
Access Tool
Targeted RNA assays for gene expression studies: custom sequencing panels with a simple workflow and automated data analysis.
These sequencing reagent kits offer increased stability and robustness over the v2 kit version.
Prepare sequencing libraries from mRNA to get a clear view of the coding transcriptome with strand-specific information.
Provides a reproducible, economical solution for sequencing RNA from FFPE tissues and other low-quality samples. Accuracy from as little as 10 ng total RNA.
Expert-defined content targeting 113 genes associated with genetic cancer risk predisposition.
Achieve rapid, targeted interrogation of an expansive number of target genes with exceptional capture efficiency and coverage uniformity.
Limited Use Label License: This product and its use are the subject of one or more issued and/or pending U.S. and foreign patent applications owned by Max Planck Gesellschaft, exclusively licensed to New England Biolabs, Inc. and sublicensed to Illumina, Inc. The purchase of this product from Illumina, Inc., its affiliates, or its authorized resellers and distributors conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product by the buyer (whether the buyer is an academic or for profit entity). The purchase of this product does not convey a license under any claims in the foregoing patents or patent applications direct to producing the product. The buyer cannot sell or otherwise transfer this product or its components to a third party or otherwise use the product for the following COMMERICAL PURPOSES: (I) use of the product or its components in manufacturing; or (2) use of the product or its components for therapeutic or prophylactic purposes in human or animals.
